Abstract
Epstein-Barr virus (EBV) is associated with a substantial number of gastric adenocarcinomas worldwide, as confirmed by EBER1/2-RNA in situ hybridization (RISH). In the present study, we developed a rapid and sensitive PCR-based prescreening method for the detection of EBV in gastric carcinomas to reduce the amount of laborious EBER1/2-RISH assays to be performed. The method was evaluated by testing gastric adenocarcinomas (n = 242) using both BamHI W PCR-enzyme immunoassay (EIA) and EBER1/2-RISH, in combination with appropriate DNA and RNA quality controls. Seventy-four percent of the paraffin-embedded gastric adenocarcinomas had good DNA quality as shown by beta-globin polymerase chain reaction (PCR) after proteinase K and boiling pretreatment, whereas after DNA purification this was increased to 90%. Thirty-two percent of all cases were EBV-DNA positive after PCR-EIA, whereas 10% of these gastric cancers contained EBV transcripts in the neoplastic cells as confirmed by EBER1/2-RISH. Interestingly, only samples with high optical density (OD) 405/630 values in PCR-EIA, equivalent to the maximum reading of the assay as determined by the positive control, contained EBV-positive tumor cells in the EBER1/2-RISH. In contrast, the weak positive samples, as determined by low OD readings in the PCR-EIA were EBER1/2-RISH negative. In conclusion, high OD values in EBV PCR-EIA are very valuable to prescreen EBV-carrying gastric carcinomas as confirmed by EBER1/2-RISH. Only these samples and those with poor DNA quality will require testing in the EBER1/2-RISH, thereby reducing the amount of laborious RISH assays with 85%.
Original language | English |
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Pages (from-to) | 870-7 |
Number of pages | 8 |
Journal | Modern Pathology |
Volume | 15 |
Issue number | 8 |
DOIs | |
Publication status | Published - Aug 2002 |
Keywords
- Actins
- Adenocarcinoma
- Comparative Study
- DNA, Viral
- Epstein-Barr Virus Infections
- Herpesvirus 4, Human
- Humans
- Immunoenzyme Techniques
- Immunohistochemistry
- In Situ Hybridization
- Journal Article
- Polymerase Chain Reaction
- RNA, Messenger
- RNA-Binding Proteins
- Research Support, Non-U.S. Gov't
- Ribosomal Proteins
- Sensitivity and Specificity
- Stomach Neoplasms
- Tumor Virus Infections